S1 domain-containing STF modulates plastid transcription and chloroplast biogenesis in Nicotiana benthamiana

New Phytol. 2012 Jan;193(2):349-63. doi: 10.1111/j.1469-8137.2011.03941.x. Epub 2011 Nov 4.

Abstract

• In this study, we examined the biochemical and physiological functions of Nicotiana benthamiana S1 domain-containing Transcription-Stimulating Factor (STF) using virus-induced gene silencing (VIGS), cosuppression, and overexpression strategies. • STF : green fluorescent protein (GFP) fusion protein colocalized with sulfite reductase (SiR), a chloroplast nucleoid-associated protein also present in the stroma. Full-length STF and its S1 domain preferentially bound to RNA, probably in a sequence-nonspecific manner. • STF silencing by VIGS or cosuppression resulted in severe leaf yellowing caused by disrupted chloroplast development. STF deficiency significantly perturbed plastid-encoded multimeric RNA polymerase (PEP)-dependent transcript accumulation. Chloroplast transcription run-on assays revealed that the transcription rate of PEP-dependent plastid genes was reduced in the STF-silenced leaves. Conversely, the exogenously added recombinant STF protein increased the transcription rate, suggesting a direct role of STF in plastid transcription. Etiolated seedlings of STF cosuppression lines showed defects in the light-triggered transition from etioplasts to chloroplasts, accompanied by reduced light-induced expression of plastid-encoded genes. • These results suggest that STF plays a critical role as an auxiliary factor of the PEP transcription complex in the regulation of plastid transcription and chloroplast biogenesis in higher plants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chloroplasts / genetics*
  • Chloroplasts / metabolism
  • Chloroplasts / ultrastructure
  • Electrophoretic Mobility Shift Assay
  • Gene Expression Regulation, Plant
  • Gene Silencing / radiation effects
  • Intracellular Membranes / metabolism
  • Intracellular Membranes / radiation effects
  • Intracellular Membranes / ultrastructure
  • Light
  • Molecular Sequence Data
  • Nicotiana / genetics*
  • Nicotiana / radiation effects
  • Nicotiana / ultrastructure
  • Phenotype
  • Photosynthesis / radiation effects
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Protein Structure, Tertiary
  • Protein Transport / radiation effects
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / metabolism
  • Subcellular Fractions / metabolism
  • Subcellular Fractions / radiation effects
  • Suppression, Genetic / radiation effects
  • Thylakoids / ultrastructure
  • Transcription, Genetic* / radiation effects

Substances

  • Plant Proteins
  • RNA, Messenger
  • RNA-Binding Proteins

Associated data

  • GENBANK/HM012811